Patient-derived tumor organoids have emerged as promising models for predicting personalized drug responses in cancer therapy,but they typically lack immune components.Preserving the in vivo association between tumor cells and endogenous immune cells is critical for accurate testing of cancer immunotherapies.Mechanical dissection of tumor specimens into tumor fragments,as opposed to enzymatic digestion into single cells,is essential for maintaining these native tumor-immune cell spatial relationships.However,conventional mechanical dissection relying on manual mincing is time-consuming and irreproducible.This study describes two microdissection devices,theμDicer andμGrater,to facilitate the generation of intact tumor fragments from mouse B16 melanoma,a common model of human melanoma.TheμDicer-andμGrater-cut tumor fragments were used to generate air-liquid interface(ALi)organoids that copreserve tumor cells with infltrating immune subsets without artificial reconstitution.TheμDicer,consisting of a hexagonal array of silicon microblades,was employed to investigate the effect of organoid size.The viability of ALI organoid immune cells appeared insensitive to organoid sizes exceeding~400μm but diminished in organoids~200μm in size.TheμGrater,consisting of an array of submillimeter holes in stainless steel,was employed to accelerate dissection.For the samples studied,theμGrater was 4.5 times faster than manual mincing.Compared with those generated by manual mincing,ALl organoids generated by theμGrater demonstrated similar viability,immune cell composition,and responses to anti-PD-1 immunotherapy.With further optimization,theμGrater holds potential for integration into clinical workflows to support the advancement of personalized cancer immunotherapy.
Microdissection testicular sperm extraction(micro-TESE)is widely used to treat nonobstructive azoospermia.However,a good prediction model is required to anticipate a successful sperm retrieval rate before performing micro-TESE.This retrospective study analyzed the clinical records of 200 nonobstructive azoospermia patients between January 2021 and December 2021.The backward method was used to perform binary logistic regression analysis and identify factors that predicted a successful micro-TESE sperm retrieval.The prediction model was constructed using acquired regression coefficients,and its predictive performance was assessed using the receiver operating characteristic curve.In all,67 patients(sperm retrieval rate:33.5%)underwent successful micro-TESE.Follicle-stimulating hormone,anti-Miillerian hormone,and inhibin B levels varied significantly between patients who underwent successful and unsuccessful micro-TESE.Binary logistic regression analysis yielded the following six predictors:anti-Mullerian hormone(odds ratio[OR]=0.902,95%confidence interval[Cl]:0.821-0.990),inhibin B(OR=1.012,95%Cl:1.001-1.024),Klinefelter’s syndrome(OR=0.022,95%Cl:0.002-0.243),Y chromosome microdeletion(OR=0.050,95%Cl:0.005-0.504),cryptorchidism with orchiopexy(OR=0.085,95%Cl:0.008-0.929),and idiopathic nonobstructive azoospermia(OR=0.031,95%Cl:0.003-0.277).The prediction model had an area under the curve of 0.720(95%Cl:0.645-0.794),sensitivity of 65.7%,specificity of 72.2%,Youden index of 0.379,and cut-off value of 0.305 overall,indicating good predictive value and accuracy.This model can assist clinicians and nonobstructive azoospermia patients in decision-making and avoiding negative micro-TESE results.
Stepwise mini-incision microdissection testicular sperm extraction(mTESE)is a procedure that attempts to minimize testicular damage.However,the mini-incision approach may vary in patients with different etiologies.Here,we performed a retrospective analysis of 665 men with nonobstructive azoospermia(NOA)who underwent stepwise mini-incision mTESE(Group 1)and 365 men who underwent standard mTESE(Group 2).The results showed that the operation time(mean±standard deviation)for patients with successful sperm retrieval in Group1(64.0±26.6min)was significantly shorter than that in Group2(80.2±31.3min),with P<0.001.The total sperm retrieval rate(SRR)was 23.1%in our study,and there was no significant difference between Group 1 and Group 2(P>0.05),even when the etiologies of NOA were taken into consideration.The results of consecutive multivariate logistic regression analysis(odds ratio[0R]:0.57;95%confidence interval[Cl]:0.38-0.87;P=0.009)and receiver operating characteristic(ROC)analysis(area under the ROC curve[AUC]=O.628)showed that preoperative anti-Mullerian hormone(AMH)level in idiopathic NOA patients was a potential predictor for surgical outcomes after initial three small incisions made in the equatorial region without sperm examined under an operating microscope(Steps 2-4).In conclusion,stepwise mini-incision mTESE is a useful technique for NOA patients,with comparable SRR,less surgical invasiveness,and shorter operation time compared with the standard approach.Low AMH levels may predict successful sperm retrieval in idiopathic patients even after a failed initial mini-incision procedure.
To investigate the factors affecting the sperm retrieval rate of microdissection testicular sperm extraction(micro-TESE)in patients with nonmosaic Klinefelter syndrome(KS),64 patients with nonmosaic KS who underwent micro-TESE in the Center for Reproductive Medicine of Peking University Third Hospital(Beijing,China)between January 2016 and December 2017 were included in the study.Data on medical history,physical examination and laboratory examination results,and micro-TESE outcomes were collected.Patients were divided into two groups according to micro-TESE outcomes.The following factors were compared between the two groups by the Mann‒Whitney U test or Student’s t-test based on the distribution(nonnormal or normal)of the factors:age,testicular size,follicle-stimulating hormone level,luteinizing hormone level,testosterone level,and anti-Müllerian hormone level.The overall success rate of sperm retrieval was 50.0%.Correlation analysis showed that testicular volume was positively correlated with testosterone level.Using a logistic regression model,age and anti-Müllerian hormone levels were found to be better predictors for the sperm retrieval rate than the other parameters.
